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(Exercise 7) Before you arrive for the Separating Organic Compounds
lab exercise, please

1. Read the lab thoroughly. Note all safety guidelines.

2. Answer these preparatory questions:

What safety procedures must you follow during this lab period?

_____________________________________________________________________

_____________________________________________________________________

_____________________________________________________________________

What property of molecules allows us to separate them by column

chromatography?

______________________________________________________________________

During column chromatography, what would happen if the frit was removed from
the chromatography column?

______________________________________________________________________

What is the purpose of the matrix? ________________________________________

What is the fractionation range of the matrix in this lab? _____________________

What is the purpose of the buffer in column chromatography? ________________

If two different compounds have the same molecular weight, could they be
separated by column chromatography? If not, why? If so, on what basis could
they be separated? _____________________________________________________

______________________________________________________________________

List the compounds that you will separate in this lab with column
chromatography:

______________________________________________________________________

What is the stationary phase in paper chromatography? _____________________

What is the mobile phase? ______________________________________________

Copyright © 2017 McGraw-Hill Education. All rights reserved. No reproduction or distribution without
the prior written consent of McGraw-Hill Education.
List the compounds that you will separate in this lab with paper chromatography:

______________________________________________________________________

Why is ninhydrin used in paper chromatography?

______________________________________________________________________

What is the formula for Rf? ______________________________________________

What does Rf signify? ____________________________________________

Calculate the Rf for the four amino-acid spots on this paper chromatogram. When
measuring the distance that each amino-acid sample traveled, measure the
highest point of each spot.

solvent front

origin
1 2 3 4

Amino acid Distance traveled by sample Distance traveled by solvent Rf

1
2
3
4

Why is important to measure each spot in a consistent way?

______________________________________________________________________

Remember that your sample moves with your solvent. If your Rf is greater than 1,
what should you do?

______________________________________________________________________

How does gel electrophoresis separate molecules? _________________________

Which molecules move the farthest on the gel? _________________________

Copyright © 2017 McGraw-Hill Education. All rights reserved. No reproduction or distribution without
the prior written consent of McGraw-Hill Education.
What is the purpose of the buffer in gel electrophoresis? _____________________

How is gel electrophoresis similar to column chromatography?

______________________________________________________________________

______________________________________________________________________

Copyright © 2017 McGraw-Hill Education. All rights reserved. No reproduction or distribution without
the prior written consent of McGraw-Hill Education.
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